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Syphilis is caused by the bacterium Treponema pallidum. The diagnosis is based on clinical data and serological analysis; however, the sensitivity and specificity of such tests may vary depending on the type of test and stage of the infection. In order to overcome this premise, this study utilized the polymerase chain reaction (PCR) for the detection of T. pallidum DNA in whole blood samples of patients with syphilis. The blood samples from patients with or without symptoms of syphilis, but with positive results in enzyme-linked immunosorbent assay (ELISA), were included in this study. A venereal disease research laboratory (VDRL) test was performed for all collected sera samples. For PCR, the T. pallidum DNA was extracted from the collected blood samples and a specific primer set was designed to amplify 131 nucleotides of polA (Tp0105). The specificity of the primers was evaluated with the DNA of 17 different pathogens. From a total of 314 blood samples reactive in ELISA, 58.2% (183/314) of the samples were reactive in the VDRL test. In the PCR, 54% (168/314) of the ELISA-reactive samples were positive. In both tests (VDRL and PCR) 104 samples were positive. Of 104 positive samples for both tests, 71 were at the latent stage. Based on these results, it can be concluded that PCR with the designed set of primers can be utilized as a diagnostic method for T. pallidum detection in blood samples of patients with syphilis, especially those with latent infection. In addition, it can be utilized as a supplement for serological methods to improve the diagnosis of syphilis.

Objective. To analyze the results of clinical and laboratory studies of children born to women with syphilitic infection, and select the most clinically significant criteria to optimize the diagnosis of early congenital syphilis. Methods. A comprehensive clinical and laboratory examination of 547 newborns born to women with a syphilitic infection. Results. The diagnostic sensitivity of the enzyme immunoassay with the determination of specific IgM was 86.6%, the polymerase chain reaction with the detection of Treponema pallidum DNA was 91.6%. Conclusion. The examination of newborns from mothers with a syphilitic infection can be optimized by reducing the number of laboratory techniques (microprecipitation reaction and passive haemagglutination reaction), subject to the mandatory determination of specific IgM and IgG in the first day of life, followed by further confirmation of diagnostics (IgG titer dynamics and the determination of the presence of T. pallidum DNA, as well as instrumental methods of research).

Background The syphilis epidemic continues to cause substantial morbidity and mortality worldwide, particularly in low- and middle-income countries, despite several recent disease control initiatives. Though our understanding of the pathogenesis of this disease and the biology of the syphilis agent, Treponema pallidum subsp. pallidum has improved over the last two decades, further research is necessary to improve clinical diagnosis and disease management protocols. Additionally, such research efforts could contribute to the identification of possible targets for the development of an effective vaccine to stem syphilis spread. Methods This study will recruit two cohorts of participants with active syphilis infection, one with de novo infection, one with repeat infection. Whole blood specimens will be collected from each study participant at baseline, 4, 12, 24, 36, and 48 weeks, to track specific markers of their immunological response, as well as to compare humoral reactivity to Treponema pallidum antigens between the two groups. Additionally, we will use serum specimens to look for unique cytokine patterns in participants with early syphilis. Oral and blood samples, as well as samples from any syphilitic lesions present, will also be collected to sequence any Treponema pallidum DNA found. Discussion By furthering our understanding of syphilis pathogenesis and human host immune response to Treponema pallidum, we will provide important data that will help in development of new point-of-care tests that could better identify active infection, leading to improved syphilis diagnosis and management. Findings could also contribute to vaccine development efforts.

Early diagnosis of Treponema pallidum infection is helpful for disease management, and conventional PCR is suitable for lesion swabs of patients with probable early syphilis. We thus tested nested and real-time PCR (NR-PCR) in various biosamples from syphilitic patients.Samples were collected from syphilis patients before treatment. Specific primer sequences targeting the T. pallidum gene polA were designed for NR-PCR.Across syphilis types, most samples assayed with NR-PCR returned a positive result, including earlobe blood (92.0%), cerebrospinal fluid (CSF) (90.2%), lesion swabs (74.3%), serum (66.9%), and whole blood (64.2%). No significant differences were observed in positive samples for whole blood, serum, and lesion swabs between primary and secondary syphilis (P0.05 for all comparisons). However, more whole-blood samples from patients with secondary syphilis were positive for NR-PCR than whole blood samples from patients with tertiary and latent syphilis (P0.05 for all comparisons). For neurosyphilis patients, significantly more earlobe blood samples tested positive than did whole-blood samples (P0.05), but there was no difference in positive results for earlobe blood and whole blood in latent syphilis. Significantly more serum samples tested positive in latent syphilis patients with rapid plasma regain (RPR) titers of 1:8 or greater, compared to those with RPR of 1:4 or less.Nested and real-time PCR can be used to identify T. pallidum DNA in biosamples from syphilitic patients, especially earlobe blood.

The aim of this work was to investigate the application of the nested PCR assay for the detection of Treponema pallidum (TP) DNA from the blood of patients with different stages of syphilis. In this study, a nested PCR method targeting the Tpp47 and polA genes (Tpp47-Tp-PCR and polA-Tp-PCR) was developed to detect TP-DNA in whole blood samples collected from 262 patients with different stages of syphilis (84 primary syphilis, 97 secondary syphilis, and 81 latent syphilis patients). The PCR assay detected T. pallidum DNA in 53.6% and 62.9% of the patients with primary and secondary syphilis, respectively, which was much higher than the detection levels in patients with latent syphilis (7.4%) (both p

Azithromycin has shown high efficacy in randomized trials when used for treating infectious syphilis in Africa. However, its use in clinical practice has been limited by the development of antimicrobial drug resistance. Resistance has not previously been reported from Australasia. The aim of this study was to determine the prevalence of and risk factors for azithromycin-resistant syphilis-causing strains in Sydney, Australia. We evaluated 409 samples that were PCR positive for Treponema pallidum DNA collected between 2004 and 2011 for the presence of the A2058G mutation, which confers resistance to macrolide antibiotics such as azithromycin. Overall, 84% of samples harbored the mutation. The prevalence of the mutation increased during the study period ( P trend, 0.003). We also collected clinical and demographic data on 220 patients from whom these samples had been collected to determine factors associated with the A2058G mutation; 97% were from men who have sex with men. Reporting sex in countries other than Australia was associated with less macrolide resistance (adjusted odds ratio, 0.25; 95% confidence interval, 0.09 to 0.66; P = 0.005), with other study factors showing no association (age, HIV status, recent macrolide use, stage of syphilis, or history of prior syphilis). Azithromycin cannot be recommended as an alternative treatment for syphilis in Sydney.

Treponema pallidum is the causative agent of syphilis, a sexually transmitted infection of significant public health importance. Since 2000 there has been a marked increase in the number of cases of syphilis infections notified in Victoria, Australia, with the majority of cases occurring in men who have sex with men (MSM) and the highest incidence being in HIV-infected MSM. The molecular subtyping method described by Pillay et al. (A. Pillay et al., Sex. Transm. Dis. 25:408–414, 1998) has been used in this study to determine the diversity of T. pallidum subtypes circulating locally and to look for any relationship between T. pallidum subtypes and HIV status over a 6-year period (2004 to 2009). Treponema pallidum DNA was detected in 303 patient specimens ( n = 3,652), and full subtyping profiles were obtained from 90 of these (from 88 patients). A total of 11 T. pallidum subtypes were identified: types 14e (28, 31.1%), 14d (15, 16.7%), 14k (13, 14.4%), 14p (12, 13.3%), 14i (7, 7.8%) 14b (6, 6.7%), 14l (5, 5.6%), and 12i, 13b, 13i, and 13e (1 each, 1.1%). This study showed a similar level of variation among circulating T. pallidum strains compared with that in other studies using the same methodology. A different mix of strains and different predominating strains have been found at each geographical study location, with type 14e emerging as the predominant local strain in Victoria. There was no detectable trend between T. pallidum subtypes and the specimen collection site or stage of syphilis (where known), nor was there any relationship between particular strains and HIV status.

The presence of Treponema pallidum DNA was assessed by real-time PCR in samples of blood donors with reactive serologic tests for syphilis. Treponema pallidum DNA was detected in two (1·02%) of 197 samples of VDRL>8, EIA+ and FTA-ABS+ donors, and in no sample from 80 VDRL−, EIA+ and FTA-ABS+ donors. Donors VDRL−, EIA+ and FTA-ABS+ lack demonstrable T. pallidum DNA in their blood and are unlike to transmit syphilis. Donors VDRL>8, EIA+ and FTA-ABS+ carry the risk of syphilis infectivity even in concomitance to antibodies detection. Serologic screening for syphilis may still play a role to prevent its transfusion transmission.

The protective response against Treponema pallidum (Tp) infection of a DNA vaccine enhanced by an adjuvant CpG ODN was investigated.The mucosal adjuvant CpG ODN enhanced the production of higher levels of anti-TpGpd antibodies induced by pcD/Gpd-IL-2 in rabbits. It also resulted in higher levels of secretion of IL-2 and IFN-γ, and facilitated T cell proliferation and differentiation (p0.05). No significant difference about testing index above-mentioned was found in the intranasal immunization group of pcD/Gpd-IL-2 vaccine adjuvanted by CpG ODN when compared with the immunization by pcD/Gpd-IL-2 vaccine intramuscular injection alone (p0.05). Furthermore, CpG ODN stimulated the production of mucosa-specific anti-sIgA antibodies and resulted in the lowest Tp-positive rate (6.7%) for Tp-infection of skin lesions and the lowest rates (8.3%) of ulceration lesions, thus achieving better protective effects.New Zealand rabbits were immunized with the eukaryotic vector encoding recombinant pcD/Gpd-IL-2 using intramuscular multi-injection or together with mucosal enhancement via a nasal route. The effect of the mucosal adjuvant CpG ODN was examined.The CpG ODN adjuvant significantly enhances the humoral and cellular immune effects of the immunization by pcD/Gpd-IL-2 with mucosal enhancement via nasal route. It also stimulates strong mucosal immune effects, thus initiating more efficient immune-protective effects.

This study aimed to reassess rates of syphilis azithromycin resistance in Dublin. Of the 104 samples, 36 (34.6%) were positive for Treponema pallidum DNA by polymerase chain reaction. Thirty samples were analyzed for A2058G mutation, 29 samples sequenced. Of the 29 samples, 27 (93.1%) had the mutation. Azithromycin cannot be recommended for the treatment of syphilis in Dublin.

Background Prior studies have reported PCR detection of Treponema pallidum DNA from whole blood and earlobe scrapings. Factors affecting efficiency such as PCR target gene and clinical stage of syphilis have been studied. We aimed to determine other factors associated with detection of T.pallidum DNA from whole blood and earlobe samples. Methods Data were obtained from baseline samples collected for a prospective cohort study of 401 men who have sex with men (MSM) and transgender women (TW) in Lima, Peru. Participants were assessed for HIV using rapid HIV testing (Alere Determine, USA), a combined Antigen/Antibody HIV EIA, and Western blot confirmation (Genscreen ULTRA HIV Ag-Ab and Genetic Systems HIV-1 Western Blot, Bio Rad, USA). Syphilis infection was assessed with RPR (BD Macro-Vue, USA) and TPPA (Fujirebio, Japan). Whole blood samples and earlobe capillary blood was collected from patients with a positive RPR and TPPA. DNA extraction was performed using the QIAamp mini kit (Qiagen, Valencia, CA), and samples were then concentrated. Ujchowsing specific primers for the TpN47 gene, an aliquot of the DNA sample was amplified using conventional PCR for participants with high RPR titers (≥1:16). Positivity was determined by visual detection of PCR product on 1% agarose gel. Results 56 participants had RPR titer ≥1:16 (1:16(n = 18), 1:32(n = 16), 1:64(n = 17), 1:128(n = 3), and 1:256(n = 2)). A total of 7 (12.5%) participants had T.pallidum DNA detected from whole blood, and 6 (10.7%) from earlobe capillary blood (10 participants total). Of these 10 participants, 5 were HIV positive, and 5 were HIV negative. Conclusion Our relatively low efficiency of detection from both whole blood and earlobe samples suggest that neither HIV status nor high RPR titer (≥1:16) seem to predict high likelihood of DNA detection. Further study is ongoing to determine if these or other characteristics are associated with positive detection of T.pallidum DNA. Disclosure of interest statement The Picasso study is funded by a grant from the United States NIAID and was implemented by the Universidad Peruana Cayetano Heredia in collaboration with the University of California, Los Angeles. The molecular part of the study was implemented under the supervision of the University of Washington. No pharmaceutical grants were received during the development of this study.

To find out if polymerase chain reaction could be used as a diagnostic tool for detecting neurosyphilis, we have applied the PCR for the detection of Treponema pallidum DNA in the cerebrospinal fluid (CSF) of syphilis patients. The results of the PCR of the CSF in 26 patients with at various stages of illness were compared with the results of other conventional tests used in the WHO criteria. T. pallidum was detected in the CSF of patients at all stages of syphilis, which indicates that they invade the central nervous system from the early stages of infection. However, the presence of T. pallidum in the CSF was not correlated with the results of other tests used in the WHO criteria, and its significance in the diagnosis of neurosyphilis should further be evaluated.